Description:

  

IsoAmpIIIUniversaltHDAkitcanbeusedtoamplifyanddetectshortDNAsequences(70bp-120bp)ataconstanttemperature.Thekitcanbeusedwithavarietyoftemplates,includingmicrobialgenomicDNA,viralDNA,plasmidDNA,andCDNA.AsinglecopyoftargetDNAcanbeamplifiedbytHDAanddetectedbyagarosegelelectrophoresiswhenoptimizedprimersandbufferareused.

IsoAmp®IIIUniversaltHDAkitisbasedonathird-generationtHDAformulaformorerobustandspecificamplificationwithreducednon-specificproductsduetoprimer-dimer.Insomecases,rapidamplificationmaybeachievedbyaddingmoreIsoAmp®IIIEnzymeMixintothereaction(unpublishedobservation).ThevolumeofIsoAmp®IIIEnzymeMixhasbeenreducedto2µLperreaction,whichallowstheusertoaddmoreenzymes(2Xor3X)forpossIBLehigh-speedamplification.IsoAmp®IIIEnzymeMix(Cat#H1020S)canbepurchasedseparately.

IsoAmp®IIIUniversaltHDAkitsupportsallreactionswhicharesupportedbyIsoAmp®IIUniversaltHDAkit.Selectedexamplesareincludedinthismanual,includingend-pointtHDAandreal-timequantitativeHDA(qHDA)inbothone-stepandtwo-stepformats.AControlTemplateandasetofspecificprimersaresuppliedforthepositivecontrolreaction.

 

  

Figure1.HDATechnology.

  

 

Figure2.PositivecontrolassayswithvaryingamountofControlTemplate.
Positivecontrolassayswereconductedfollowingtheone-stepqHDAprotocol.ControlTemplatewasusedat5ng-500fgperassayintriplicate.

*Negativecontrolreactionsmayproduceprimer-dimer,whichcanbedistingushedfromspecificampliconsbygel-analysisorbymelting-curveanalysis.

 

Applications:

  

UsedtoamplifyanddetectshortDNAsequences(80-120bp)

-downloadManual

Advantages:
UseofhelicasetoseparateDNAeliminatesneedforThermocycler
Reactionscanbeperformedatconstanttemperature.
Workswithavarietyoftemplates,includingbacterialgenomicDNA,viralDNA,plasmidDNAandcDNA.
AslowasasinglecopyoftargetDNAcanbeamplifiedunderoptimizedconditions.
Canbeusedforreal-timequantitativeor/andreversetranscriptionHDA.

   

KitComponents:

  

IsoAmp®IIIEnzymeMix(50reactions)
IsoAmp®dNTPSolution(50reactions)
10XAnnealingBufferII
MgSO₄(100mM)
NaCl(500mM)
pCNG1(ControlTemplate:1ng/µl)
NGF3(ForwardPrimer:5µM)
NGR3(ReversePrimer:5µM)
     

StorageConditions:

  

StorageTemperature:-20ºC

RecommendedstorageconditionsforthetHDAkitare-20°Cforshort-term(lessthan6months)storageand-80°Cforstoragegreaterthan6months.     

QualityControl:

 

QualityAssuranceStatement:
KitcomponentswereindividuallytestedforoptimalperformanceintHDA.

     

Legal:

 
Licenses/Patents/Disclaimers:
NOTICETOPURCHASER:ThisproductiscoveredbyU.S.patents7,282,328and7,662,594andpendingforeignpatents.Thepurchasepriceofthisproductincludesalimited,non-transferablelicenseforresearchuseonlyunderthepatentsownedbyBiohelix.Nootherlicenseunderthesepatentsisconveyedexpresslyorbyimplicationtothepurchaserbythepurchaseoftheproduct.TheProductisnottobeusedfordiagnosticpurposesnorisitintendedforhumanuse.Thisproductmaynotberesold,modifiedforresale,orusedtomanufacturecommercialproductswithoutwrittenapprovalofBioHelixCorp.FurtherinformationregardingthisproductmaybeobtainedbycontactingBioHelixatinformation@biohelix.com.

   

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